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羊膜与脂肪来源干细胞共培养体系增强大鼠牙周缺损模型中的骨再生

Amniotic membrane and adipose-derived stem cell co-culture system enhances bone regeneration in a rat periodontal defect model.

作者信息

Wu Pin-Hsien, Chung Hsiang-Yi, Wang Jyh-Horng, Shih Jin-Chung, Kuo Mark Yen-Ping, Chang Po-Chun, Huang Yao-Der, Wang Peng-Cheng, Chang Cheng-Chi

机构信息

Graduate Institute of Oral Biology, School of Dentistry, National Taiwan University, Taipei, Taiwan.

Graduate Institute of Clinical Dentistry, School of Dentistry, National Taiwan University, Taipei, Taiwan.

出版信息

J Formos Med Assoc. 2016 Mar;115(3):186-94. doi: 10.1016/j.jfma.2015.02.002.

Abstract

BACKGROUND/PURPOSE: Periodontal disease is a chronic inflammatory process that potentially leads to alveolar bone destruction and tooth loss. Tissue engineering combined with stem cell therapy is a potential effective treatment for periodontal bone loss. Amniotic membrane (AM) is a potential scaffold enriched with multiple growth factors. It has the effects of anti-inflammation, antiangiogenesis, and immunosuppression. Herein, we used adipose-derived stem cells (ADSCs) and an AM co-cultured system to study bone regeneration in a rat periodontal defect model in vivo.

METHODS

Human ADSCs were isolated from the infrapatellar fat pad, and characterized by flow cytometry, reverse transcription-polymerase chain reaction, and multipotent differentiation assays. The co-culture system was applied in the periodontal two-wall osseous defect in a rat model, and computed tomography was used to measure the effect.

RESULTS

Human ADSCs isolated from the infrapatellar fat pad showed spindle-like morphology. Flow cytometry results demonstrated that ADSCs expressed a high level of CD90 and CD105, but not CD31, CD34, and CD45. ADSCs strongly expressed stemness genes, including SOX2, OCT4, NANOG, and KLF4 on different passages. Furthermore, ADSCs were able to differentiate into osteogenic, chondrogenic, and adipogenic cells. In the periodontal osseous defect rat model, ADSCs and the AM co-culture system significantly increased bone regeneration.

CONCLUSION

This study provides the basis for using ADSCs with an AM co-culture system as stem cell therapy and scaffold transplantation in clinical periodontology.

摘要

背景/目的:牙周病是一种慢性炎症过程,可能导致牙槽骨破坏和牙齿脱落。组织工程与干细胞治疗相结合是治疗牙周骨丧失的一种潜在有效方法。羊膜(AM)是一种富含多种生长因子的潜在支架材料。它具有抗炎、抗血管生成和免疫抑制作用。在此,我们使用脂肪来源干细胞(ADSCs)与羊膜共培养系统,在大鼠牙周缺损模型中研究体内骨再生情况。

方法

从髌下脂肪垫分离人ADSCs,并通过流式细胞术、逆转录-聚合酶链反应和多能分化试验进行鉴定。将共培养系统应用于大鼠模型的牙周双壁骨缺损,并使用计算机断层扫描测量效果。

结果

从髌下脂肪垫分离得到的人ADSCs呈纺锤样形态。流式细胞术结果表明,ADSCs高表达CD90和CD105,但不表达CD31、CD34和CD45。ADSCs在不同传代时均强烈表达干性基因,包括SOX2、OCT4、NANOG和KLF4。此外,ADSCs能够分化为成骨细胞、软骨细胞和脂肪细胞。在牙周骨缺损大鼠模型中,ADSCs与羊膜共培养系统显著促进了骨再生。

结论

本研究为在临床牙周病学中使用ADSCs与羊膜共培养系统进行干细胞治疗和支架移植提供了依据。

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